Reference : Mass-spectrometry-based method for screening of new peptide ligands for G-protein-cou...
Scientific journals : Article
Physical, chemical, mathematical & earth Sciences : Chemistry
http://hdl.handle.net/2268/182082
Mass-spectrometry-based method for screening of new peptide ligands for G-protein-coupled receptors
English
Cologna Takeno, Camila mailto [Université de Liège - ULg > Département de chimie (sciences) > Laboratoire de Spectrométrie de Masse (L.S.M.) > >]
Gilles, Nicolas mailto [Commissariat à l'Energie Atomique (Saclay) - CEA > DSV/iBiTec-S/SIMOPRO > Toxins, Receptors and Channels team > >]
Echterbille, Julien mailto [Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.) > >]
Degueldre, Michel mailto [Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.) > >]
Servent, Denis mailto [Commissariat à l'Energie Atomique (Saclay) - CEA > DSV/iBiTec-S/SIMOPRO > Toxins, Receptors and Channels team > >]
De Pauw, Edwin mailto [Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.) > >]
Quinton, Loïc mailto [Université de Liège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M) >Chimie biologique > > >]
2015
Analytical and Bioanalytical Chemistry
Springer Science & Business Media B.V.
407
5299-5307
Yes (verified by ORBi)
International
1618-2642
1618-2650
Berlin
Germany
[en] Venomics ; mass spectrometry ; screening ; GPCR ; peptide ligand
[en] G-protein-coupled receptors (GPCRs) constitute the largest family of transmembrane proteins. Although implicated in almost all physiological processes in the human body, most of them remain unexploited, mostly because of the lack of specific ligands. The objective of this work is to develop a new mass-spectrometry-based technique capable of identifying new peptide ligands for GPCRs. The strategy is based on the incubation of cellular membranes overexpressing GPCRs with a mixture of peptides that contains potential ligands. Peptide ligands bind to the receptors, whereas other peptides remain in the binding buffer. Bound peptides are eluted from membranes and directly detected, identified, and characterized by MALDI TOF–TOF. The results reveal the efficacy of the procedure for selecting a specific ligand of GPCRs in both simple and complex mixtures of peptides. This new approach may offer direct purification, identification, and characterization of the new ligand in a single workflow. The proposed method is labeling-free and, unlike radio-binding and other techniques, it does not require a previously known labeled ligand of the studied GPCR. All these properties greatly reduce the experimental constraints. Moreover, because it is not based on the principle of a competitive specific binding, this technique constitutes a new tool to discover new ligands not only for known GPCRs, but also for orphan GPCRs
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/182082
10.1007/s00216-015-8692-4
http://link.springer.com/article/10.1007%2Fs00216-015-8692-4#page-1

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